Feed supplement and a feed composition comprising resin acid based composition

ABSTRACT

The present invention relates to a feed supplement which comprises a resin acid based composition comprising over 10% (w/w) resin acids for use in the prevention of growth of harmful bacteria in the animal digestive tract, in the prevention of intestinal disorders, in the modulation of microbial population of the animal digestive tract, in enhancing rumen fermentation, lowering rumen methane production and/or in binding toxins. The invention further relates to a use of the feed supplement and a feed composition comprising the feed supplement.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a Continuation of International ApplicationPCT/FI2014/050832, filed Nov. 5, 2014, which international applicationwas published on May 21, 2015, as International PublicationWO2015/071534. The International Application claims priority of FinnishPatent Application 20136113, filed Nov. 13, 2013, the contents of whichare incorporated herein by reference in their entireties.

FIELD OF THE INVENTION

The invention relates to a feed supplement and a feed compositioncomprising resin acid based composition and to an use of the feedsupplement.

BACKGROUND OF THE INVENTION

Imbalances in microbial populations and growth of harmful bacteria inthe digestive tract of animals can cause significant losses in animalgrowth and production. These imbalances manifest themselves asintestinal disorders such as diarrhea. While microbial infections ofanimals have been prevented by the use of e.g. antibiotics and otheragents that prevent the growth of microorganisms, stricter regulationson their use are expected. Ruminant animals can utilize fiber-rich rawmaterials which have little or no nutritional value for monogastricslike the human. However, the feed conversion efficiency of ruminants isrelatively low and their methane production represents a remarkableshare of the world's greenhouse gas emissions. With the increasingdemand of food there is a need to improve the feed conversion efficiencyof ruminants and to lower their methane production. Generally, there isan increasing demand for ingredients for use in animal feeding that canmodulate the microbial population in the animal digestive tract butwhich are readily available, well tolerated and environmentallyfriendly.

Fractional distillation of crude tall oil (CTO), obtained as aby-product of the Kraft process of wood pulp manufacture, producesdepitched tall oil which typically comprises over 10% resin acids andless than 90% fatty acids. Further refinement of depitched tall oilproduces tall oil fatty acid (TOFA), Distilled Tall Oil (DTO) and TallOil Rosin (TOR) which are available in a variety of compositionsdiffering in the fatty acids and resin acids content. Because TOFA is aninexpensive source of fatty acids, it has previously been used in animalnutrition as an energy source. For instance, GB 955316 discloses the useof alkali metal salts of tall oil fatty acids to improve weight gain andnitrogen retention in ruminant animals.

Toxins are poisonous substances produced within living cells ororganisms. Toxins such as mycotoxins are a chemically variable group ofsecondary metabolites of fungi, which can be found in grains and otherfeedstuffs even in the absence of any visible fungal growth. Hightemperature and air humidity during the storage increase the likelihoodof fungal growth, but mycotoxin contamination can also occur already inthe field. Visible appearance or smell of grains or silage does notindicate the presence or absence of mycotoxin contamination. Effects oftoxins such as mycotoxins to farm animals are very variable, and rangefrom increased mortality to decreased fertility and performance.Mycotoxins may also disturb the immune system of animals and make themmore susceptible to diseases.

Due to the chemical variability of mycotoxins, analysis of all feedlotsfor even the most common mycotoxins would be too expensive. Thereforemycotoxin adsorbents are often used to give extra insurance againstmycotoxin contamination in feeds. Mycotoxin adsorbents are substancesthat are itself not digested or absorbed by the animal. They are assumedto bind toxins during the passage through the alimentary canal. Thus,instead of being absorbed by the animals, the toxins get eventuallyvoided via feces.

Toxin binders can also adsorb other types of toxins, like bacterialtoxins or secondary metabolites of plants from the digestive tract.Activated carbon (charcoal) is an efficient toxin binder. It is arecommended general toxin binder in various poisonings. However,charcoal also binds vitamins and minerals, which makes it unsuitable forcontinuous use in feeds.

PURPOSE OF THE INVENTION

The purpose of the invention is to provide a new type of feed supplementcomprising resin acid based composition for use in the prevention ofgrowth of harmful bacteria in the animal digestive tract, in theprevention of intestinal disorders, in the modulation of microbialpopulation of the animal digestive tract, in enhancing rumenfermentation, lowering rumen methane production and/or in bindingtoxins.

SUMMARY

The feed supplement according to the present invention is characterizedin that said feed supplement comprises the resin acid based compositioncomprising over 10% (w/w) resin acids for use in the prevention ofgrowth of harmful bacteria and/or in the prevention of intestinaldisorders and wherein the resin acid based composition is Tall Oil Rosin(TOR), Wood Rosin, GUM Rosin and/or Distilled Tall Oil (DTO).

The use of the feed supplement according to the present invention ischaracterized by using a feed supplement comprising the resin acid basedcomposition comprising over 10% (w/w) resin acids in the modulation ofmicrobial population of the animal digestive tract by increasing theconcentrations of acetic and propionic acids and decreasing theconcentration of lactic acid, and for improving feed utilization.

The use of the feed supplement according to the present invention ischaracterized by using a feed supplement comprising the resin acid basedcomposition comprising over 10% (w/w) resin acids in binding toxins.

The use of the feed supplement according to the present invention ischaracterized in that the toxin is mycotoxin.

The feed composition according to the present invention is characterizedin that it comprises a feed supplement in an amount of 0.00001-1.0%(w/w) of the dry weight of the total amount of feed and is used in theprevention of growth of harmful bacteria and/or in the prevention ofintestinal disorders.

The use of the feed supplement according to the present invention ischaracterized in that the feed composition comprises a feed supplementin an amount of 0.00001-1.0% (w/w) of the dry weight of the total amountof feed and is used in the modulation of microbial population of theanimal digestive tract by increasing the concentrations of acetic andpropionic acids and decreasing the concentration of lactic acid, inenhancing rumen fermentation, lowering rumen methane production and/orin binding toxins.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the turbidity change during 8 hours of Cl. perfringensgrowth as a response to a Tall Oil Rosin (TOR) and Distilled Tall Oil(DTO).

FIG. 2 shows gas production during 8 hours by Cl. perfringens growth asa response to Tall Oil Rosin (TOR) and Distilled Tall Oil (DTO).

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based on the realization that a feed supplementwhich comprises resin acid based composition can be used in theprevention of growth of harmful bacteria in the animal digestive tract,in the modulation of microbial population of the animal digestive tract,in the prevention of intestinal disorders, in enhancing rumenfermentation, lowering rumen methane production and/or in bindingtoxins.

Resin acids are present in coniferous trees, and there are three mainspecies of resin acid products, namely Tall Oil Rosin (TOR), Wood Rosinand GUM Rosin. TOR is the resin acid fraction separated by vacuumdistillation from Crude Tall Oil (CTO) which is produced by thepreparation of pulp. CTO is obtained via acidulation of Crude Tall OilSoap or Crude Sulphate Soap (TOS). TOS is separated from cooking liquidin pulp mill often called black liqueur during pulping process. WoodRosin is the fraction separated by steam distillation or other meansfrom dead trees, tree stumps, branches etc. and GUM Rosin is the resinfraction that has been steam distilled or separated by other means fromresin harvested often called tapping from a living tree.

GUM resin is widely produced in China, Indonesia and Brazil. Wood rosinmainly comes from the USA. TOR is produced in the USA and Scandinaviaand to a lesser extent in Central Europe, New Zealand and Russia.Substances containing resin acid and obtained by vacuum distillationfrom crude tall oil include Distilled Tall Oil (DTO), Tall Oil FattyAcid (TOFA) and Tall Oil Pitch (TOP). DTO contains 10-40% resin acids.CTO typically contains 15-70% resin acids, and the lowest resin acidcontents are generally provided by the cooking of mixed wood pulp.

The term “Tall Oil Rosin” or “TOR” should be understood as referring toa composition obtained by distillation of crude tall oil and furtherrefinement of distilled tall oil. TOR typically comprises 60-99% (w/w)resin acids.

The term “Wood Rosin” should be understood as referring to a compositionobtained by distillation or other means from dead trees, tree stumps,branches etc. Wood Rosin typically comprises 50-99% (w/w) resin acids.

The term “GUM Rosin” should be understood as referring to a compositionobtained by distillation or separated by other means from resinharvested from a living tree. GUM Rosin typically comprises 50-99% (w/w)resin acids.

The term “Distilled Tall Oil” or “DTO” should be understood as referringto a composition obtained by distillation of crude tall oil and furtherrefinement of distilled tall oil. DTO typically comprises 10-60% (w/w)resin acids.

The resin acid based composition TOR, Wood Rosin, GUM Rosin, CTO, TOSand DTO can also be produced by mixing one or more resin acidcompositions and one or more fatty acid compositions in form of oils orfats. Produced resin acid derivatives are for example esters, ethers oralkali metal salts.

Resin acids are known to have antimicrobial, including antibacterial,properties.

The feed supplement of the present invention comprises a resin acidbased composition which comprises over 10% (w/w) of resin acids.

In one embodiment of the present invention, the feed supplementcomprises a resin acid based composition which comprises over 12% (w/w)resin acids.

In one embodiment of the present invention, the feed supplement iseffective in the prevention of growth of harmful bacteria, forprevention of intestinal disorders, in the modulation of microbialpopulation of the animal digestive tract, in enhancing rumenfermentation, lowering rumen methane production and/or in bindingtoxins.

In one embodiment of the present invention, the feed supplementcomprising a resin acid based composition is for use in the preventionof growth of harmful bacteria and/or in the prevention of intestinaldisorders.

In one embodiment of the present invention, the feed supplementcomprising a resin acid based composition is used in the modulation ofmicrobial population of the animal digestive tract.

In one embodiment the feed supplement is used for improving feedutilization. In one embodiment the feed supplement is used for improvingthe feed conversion ratio.

In one embodiment of the present invention, the feed supplementcomprising a resin acid based composition is used in enhancing rumenfermentation and/or lowering rumen methane production.

In one embodiment of the present invention, the feed supplementcomprising a resin acid based composition is used in binding toxins.

In one embodiment, the feed supplement comprising a resin acid basedcomposition is used in binding mycotoxins.

In this context, the term “feed supplement” should be understood asreferring to a composition that may be added to a feed or used as suchin the feeding of animals.

In this context, the term “resin acids” should be understood asreferring to a complex mixture of various acidic compounds derived fromwood, specially pine wood. They can also be modified resin acids such asdimers and decarboxylated resin acids. The exact composition of theresin acids present in the resin acid based composition varies e.g.according to the species of the trees the composition is obtained fromand the processing conditions under which it is manufactured. Resinacids typically include compounds such as abietic acid, dehydroabieticacid, levopimaric acid, neoabietic acid, pimaric acid and isopimaricacid, only to mention a few.

In the context of the feed additive, the resin acid based compositionmay be any composition described in this specification.

In one embodiment of the present invention the resin acid basedcomposition of the feed supplement comprises at least one of followingresin acids abietic acid, dehydoabietic acid, palustric acid, neoabieticacid, pimaric acid and isopimaric acid and/or derivatives thereof. Thederivatives are obtained by modifying the resin acid chemically,biologically or other ways. In one embodiment of the present inventionthe resin acid based composition comprises at least one of followingresin acids abietic acid, dehydoabietic acid, palustric acid, neoabieticacid, pimaric acid and isopimaric acid. In one embodiment of the presentinvention the resin acid based composition comprises at least onechemically modified resin acid of abietic acid, dehydoabietic acid,palustric acid, neoabietic acid, pimaric acid and isopimaric acid. Theresin acid based composition may also be a mixture of unmodified andmodified resin acids.

In one embodiment of the present invention the resin acid basedcomposition is Tall Oil Rosin (TOR).

In one embodiment of the present invention the resin acid basedcomposition and/or TOR comprises over 60% (w/w) resin acids. In oneembodiment of the present invention the resin acid based compositionand/or TOR comprises over 85% (w/w) resin acids.

The TOR can comprise 32-44.5% abietic acid, 18-25% dehydoabietic acid,0-3% dihydoabietic acid, 3.0-11.5% isopimaric acid, 0-1.5%8,5-isopimaric acid, 0-2.5% levopimaric acid, 3.3-4% neobietic acid,7.5-10% palustric acid, 3-4.5% pimaric acid and 0-4.0% sandaropimaricacid. TOR may comprise <0.1% dimers and 0-7% other components.

In one embodiment of the present invention the resin acid basedcomposition is Wood Rosin.

In one embodiment of the present invention the resin acid basedcomposition and/or Wood Rosin comprises over 10 and up to 99% (w/w)resin acids. In one embodiment of the present invention the resin acidbased composition and/or Wood Rosin comprises 50-99% (w/w) resin acids.

The Wood Rosin can comprise 45-51% abietic acid, 7.9-8.5% dehydoabieticacid, 0-1% dihydoabietic acid, 11-15.5% isopimaric acid, 0-4.2%8,5-isopimaric acid, 0-0.2% levopimaric acid, 4.7-7% neobietic acid,8.2-10% palustric acid, 3-7.1% pimaric acid and 0-2.0% sandaropimaricacid. Wood Rosin may comprise 0-4.2% dimers and 0-1% other components.

In one embodiment of the present invention the resin acid basedcomposition is GUM Rosin.

In one embodiment of the present invention the resin acid basedcomposition and/or GUM Rosin comprises over 10 and up to 99% (w/w) resinacids. In one embodiment of the present invention the resin acid basedcomposition and/or GUM Rosin comprises 50-99% (w/w) resin acids.

The GUM Rosin can comprise 15-45% abietic acid, 3-15% dehydoabieticacid, 0-0.6% dihydoabietic acid, 3.6-28% isopimaric acid, 0-0.3%8,5-isopimaric acid, 0-1.8% levopimaric acid, 10-19% neobietic acid,5-25% palustric acid, 2-7.4% pimaric acid and 0-1.5% sandaropimaricacid. GUM Rosin may comprise 0-1.0% dimers and 0-3.5% other components.

In one embodiment of the present invention the resin acid basedcomposition is Distilled Tall Oil (DTO). In one embodiment of thepresent invention the resin acid based composition is a distillationfraction of Tall Oil. In one embodiment of the present invention theresin acid based composition is a mixture of DTO and a distillationfraction of Tall Oil.

The Distillation fraction of Tall Oil is any resin acids containingfraction of CTO available during CTO refining.

In one embodiment of the present invention the resin acid basedcomposition and/or DTO comprises over 10 and up to 60% (w/w) resinacids. In one embodiment of the present invention the resin acid basedcomposition and/or DTO comprises over 10 and up to 40% (w/w) resinacids.

In one embodiment of the present invention the resin acid basedcomposition is separated from black liqueur during pulping process orTOS or CTO.

The resin acids of the resin acid based composition are insoluble inwater. The resin acids of the resin acid based composition may beunmodified or modified.

In one embodiment of the present invention the resin acids of the resinacid based composition and the feed supplement are unmodified. The term“unmodified” should be understood as referring to the resin acid basedcomposition comprising over 10% (w/w) resin acids that is not modified,i.e. treated chemically, or biologically. The feed supplement comprisingthe resin acid based composition may be used as such.

In one embodiment of the present invention the resin acids of the resinacid based composition are chemically, biologically or other waysmodified resin acid compositions. The chemical and/or biologicalmodification of resin acids of the resin acid based composition improvesthe solubility of its components and resin acids in the digestive tractof an animal. The resin acid based composition may be chemicallymodified e.g. partially or totally hydrogenated, disportinated,isomerized, oxidized, polymerized, etherified, saponified and/oresterified with suitable compounds, for example, fatty alcohols, glycol,glycerol or glyceridic fatty acid compounds such as mono- di- and tri-and polyglycerides or sugar or polyol based esters. They may be alsoused as a reactant in Diels-Alder reaction.

In one embodiment of the present invention, the feed supplementcomprises a resin acid based composition which is modified bysaponification.

Various processes for the saponification of the resin acid basedcomposition using e.g. NaOH or CaOH are known to a person skilled in theart. In one embodiment of the present invention, the resin acid basedcomposition for use according to the present invention is modified byetherification.

In one embodiment of the present invention the resin acid basedcomposition of the feed supplement comprises 1-90 (w/w) fatty acidsand/or their derivatives. The fatty acids may be in form of oils or fatsor in other forms like free fatty acids or esters, ethers or alkalimetal salts or fatty alcohols.

In one embodiment of the present invention, the resin acid basedcomposition includes unsaponifiables which have not an acid group, forexample, lipophilic neutral substances and esters from wood. In oneembodiment of the present invention, the resin acid based compositionincludes less than 1% unsaponifiables. The amount of unsaponifiables istypically in DTO products less than 5% and in TOR, Wood and GUM Rosinless than 6%.

In one embodiment of the present invention, the feed supplementcomprises resin acid based composition which is dried. The resin acidbased composition can be dried by spray drying, drum drying or by anyother known suitable drying method.

In one embodiment of the present invention, the feed supplementcomprises different active ingredients.

The feed supplement may be added in the feed in a concentration of0.0001-10 kg//ton of dry weight of the total amount of the feed. Thefeed supplement comprising the resin acid based composition according tothe invention may be added to the feed or feed supplement as such, or itmay in general be further processed as desired.

The feed supplement comprising resin acid based composition according tothe present invention can be modified into a form which is functionaland effective in feeds. Carriers such as oil, fatty acids can be addedto the composition for improving the functionality. Further emulgatorssuch as glycerols, lecithin etc. can be added to the resin acid basedcomposition for improving the solubility.

In one embodiments the feed supplement comprising the resin acid basedcomposition according to the present invention may comprise chemicallymodified resin acid derivatives The resin acid derivatives could alsocomprise rosin based acid anhydrides, dimers, amines, maleimides,alkenyls, epoxy compositions and/or mixtures thereof or with othersuitable chemically modified resin acids known to person skilled in theart.

In one embodiment of the present invention, the feed supplementcomprises resin acid based composition which is absorbed into a carriermaterial suitable for the feed composition such as sugarbeet pulp.

In one embodiment of the present invention, the feed supplementcomprises resin acid based composition which is mixed with a liquidcarrier material suitable for the feed composition such as vegetableoils or fatty acids.

Further, the feed supplement comprising the resin acid based compositionaccording to the invention may be added to the feed, or it may beadministered to an animal separately (i.e. not as a part of any feedcomposition).

In this context, the term “feed composition” or “feed” should beunderstood as referring to the total feed composition of an animal dietor to a part thereof, including e.g. supplemental feed, premixes andother feed compositions. The feed may comprise different activeingredients.

The present invention also relates to a feed composition comprising thefeed supplement according to the invention.

In one embodiment of the present invention, the feed compositioncomprises the feed supplement in an amount of 0.00001-1.0% (w/w of thedry weight of the total amount of the feed.

The present invention also relates to a use of the feed supplementaccording the present invention in a feed composition.

The invention also relates to a method of preventing the growth ofharmful bacteria in the animal digestive tract, comprising the step ofadministering to an animal the feed supplement comprising the resin acidbased composition according to the invention.

The invention also relates to a use of the feed supplement comprisingthe resin acid based composition comprising over 10% (w/w) resin acidsin modulating microbial population of the animal digestive tract,preventing intestinal disorders, in enhancing rumen fermentation,lowering rumen methane production and/or binding toxins.

In this context, the term “harmful bacteria” should be understood asreferring to any bacteria that is capable of affecting the digestivetract or health of an animal in an adverse manner, including competitionfor nutrients with the host animal. In this context, the term “microbialpopulation” should be understood as referring to the microorganisms thatinhabit the digestive tract, including the Bacteria and Archaea domainsand microscopic members of the Eukaryote domain and also intestinalparasites. The microbial population will vary for different animalspecies depending on e.g. the health of an animal and on environmentalfactors.

In this context, the term “intestinal disorder” should be understood asreferring to various disorders of the digestive tract in an animal,including e.g. diarrhea and other intestinal health problems.

In this context, the term “animal” should be understood as referring toall kinds of different animals, such as monogastric animals, ruminants,fur animals, pets and aquaculture. Non-limiting examples of differentanimals, including offspring, are cows, beef cattle, pigs, poultry,sheep, goats, horses, foxes, dogs, cats and fish.

In this context, the term “toxin” should be understood as referring toany poisonous substance produced within living cells or organisms.Toxins are products of plants, animals, microorganisms, for examplebacteria, viruses, fungi, rickettsiae, protozoa, etc. In this context,the term “mycotoxin” should be understood as referring to a toxicsecondary metabolite produced by fungi, such as yeast and mould. Themost common mycotoxins in grains or silage are for example aflatoxins,zearalenone, ochratoxin A, deoxynivalenol, fumonisin and T-2 toxin. Thetoxins will vary depending on environmental factors.

In one embodiment of the present invention, the resin acid basedcomposition is administered to an animal in an effective amount.

The feed supplement comprising the resin acid based compositioncomprising over 10% (w/w) resin acids is effective in the prevention ofgrowth of harmful bacteria in the animal digestive tract, in theprevention of intestinal disorders, in the modulation of microbialpopulation of the animal digestive tract, in enhancing rumenfermentation, lowering rumen methane production and/or in bindingtoxins. They have potential in toxin binding.

The present invention has a number of advantages. The feed supplementcomprising the resin acid based composition is a readily available,natural, low-cost and environmentally friendly material. Further, it isnon-toxic and well tolerated. The feed supplement comprising the resinacid based composition can be used as such. The invention is effectivein modulating the composition of the microbiota in the animal digestivetract to a direction that is beneficial for animal performance.Subsequently, other benefits of the invention are e.g. improved animalhealth and productivity, higher product quality, uniformity, nutritionalvalue and food and product safety, lower costs per production unit anddecreased environmental loads. The invention allows the production offeed compositions and supplements at low cost.

The embodiments of the invention described hereinbefore may be used inany combination with each other. Several of the embodiments may becombined together to form a further embodiment of the invention. Aproduct, a method or a use, to which the invention is related, maycomprise at least one of the embodiments of the invention describedhereinbefore.

EXAMPLES

In the following, the present invention will be described in moredetail.

Example 1 Pathogen Inhibition Test

Clostridium perfringens is a pathogenic bacterium that causes necroticenteritis in broiler chicks and other species of poultry. Thisexperiment was conducted to study the inhibition of Cl. perfringens bythe resin acid based compositions.

Two resin acid based compositions Tall Oil Rosin (TOR) and DistilledTall Oil (DTO) obtained from Crude Tall Oil distillation were tested astheir efficiency against Clostridium perfringens growth. The TORcomposition contained 88% (w/w) resin acids and the DTO compositioncontained 27.5% (w/w) resin acids.

Test compounds TOR (free resin acids 88%) 0.03 g of 1:1 in turnip rapeoil DTO (free resin acids 27.5%) 0.015 g TOR (free resin acids 88%) 0.15ml of 10% stock solution in ethanol DTO (free resin acids 27.5%) 0.15 mlof 10% stock solution in ethanol TOR (free resin acids 88%) 0.15 ml of1% stock solution in ethanol DTO (free resin acids 27.5%) 0.15 ml of 1%stock solution in ethanol ethanol 0.15 ml ethanol

The efficiency of test compositions was tested in a Cl. perfringensgrowth inhibition test that measures both the turbidity of theclostridial culture medium as a result of increased number of bacterialcells in a unit volume of medium, and the cumulative gas productionduring the simulation.

The efficiency of TOR and DTO against the growth of Cl. perfringens wastested at concentrations 0.01%. The TOR with 88% resin acid was meltedat +105° C. and mixed 1:1 in turnip rape oil, in order to achieve thesame runny form as the other two oily products. This diluted product wasdosed as double amount in the simulation.

Simulation Procedure:

The simulation was conducted in 25-ml glass bottles containing 15 ml ofsterile anaerobic TSGY-media (tryptic soy broth-yeast extract media withglucose) and the bottles were enclosed with air-tight stoppers to ensureanaerobic conditions throughout the experiment. At the beginning of thesimulation 0.1% inoculums of the overnight grown Cl. perfringens culturewas injected to TSGY-bottles. Test compounds, or sterile deionized waterfor the control treatment, were added in a 150 μl final volume from therespective stock solution according to the treatment. The simulationbottles were randomized to avoid artificial bias between treatments. Thebottles were kept at an even 37° C. temperature and mixed 1 min beforethe turbidity measurement at each time point. The total simulation timewas 8 h.

The optical density was measured at the time points of 0.5, 4 and 8hours. The turbidity (optical density, OD) of growth media increasesproportionally as the Cl. perfringens cell number and cell densityincreases.

The total gas production was measured at the end of the 8 h simulationas an indicator of growth efficiency, since Cl. perfringens produces gasdue to the active metabolism during exponential growth.

Results

The results are illustrated in FIGS. 1 and 2. The TOR and DTO treatmentsvery effectively inhibited the growth of Cl. perfringens, which wasdetected as the lack of turbidity change (FIG. 1) and the production ofnegligible amounts of gas (FIG. 2). TOR and DTO compositions inhibitedthe growth of Clostridium perfringens very efficiently regardless of therosin acid concentration

Example 2 Methane Inhibition Test

Two resin acid based compositions Tall Oil Rosin (TOR) and DistilledTall Oil (DTO) were tested in methane inhibition test. The TORcomposition contained 88% (w/w) resin acids and the DTO compositioncontained 27.5% (w/w) resin acids. The TOR composition containing 88%(w/w) resin acids was mixed 1:1 with turnip rape oil.

The methane inhibition test was conducted with rumen-fistulated dairycows in order to study the potential of TOR and DTO to decrease the rateof methane production in the rumen. Rumen fluid samples were measuredfor the numbers of methanogenic bacteria, as they are themethane-producing organisms. The short chain fatty acid profiles,including the concentration of branched chain fatty acids, of thesamples were measured as they indicate whether resin based acidcompositions had effects to ruminal fermentation.

Three rumen-fistulated, lactating dairy cows were given 3.0 g of drytest compositions/head/day for 21 days, in four portions. TOR and DTOcompositions were first dried onto sugar beet pulp and then mixed intothe compound feed. Rumen samples were taken before the dietaryintervention, once a week during the test composition feeding, and aftera two-week washout period. The samples of the trial were analysed forshort chain fatty acids (SCFAs) by gas chromatography and numbers ofmethanogens, protozoa and total bacteria by qPCR.

Results

The results show that the numbers of methane producing bacteriadecreased numerically during the TOR and DTO feeding period, whileprotozoa and the total number of bacteria were not affected by theproduct. The levels of lactic, propionic, and valeric acids and totalshort chain fatty acids tended to decrease in the rumen fluid during theTOR and the DTO feeding period. The concentration and relativeproportion of branched chain fatty acids tended to decrease as aresponse to TOR and the DTO.

The experiment shows that the TOR and the DTO lowers the amount ofmethanogens and thus lowers rumen methane production. The experimentalso shows that the TOR and the DTO enhances rumen fermentation.

Example 3

This experiment was conducted to study the effect of saponified DTO with35% (w/w) resin acids with or without Sugar Beet Pulp (SBP) carrier onthe microbial microbial population and fermentation of broiler chickileum in vitro.

The saponified DTO was manufactured by adding NaOH (sodium hydroxide) toDTO, adding enough water to adjust the total dry matter (DTO) percentageof the mixture to 18-20%, heating the mixture to +90° C., keeping thetemperature at +90° C. for 120 minutes, during which time the mixturewas gently stirred at 15 min intervals.

Experiment

Ileal contents of 40-days old broiler chicks were used for thesimulation media and as inoculants in the simulation models. The trialtreatments were prepared from a batch of saponified DTO.

Preparations of DTO with 35% resin acids were produced:

-   -   1. Saponified DTO with 20% dry matter content        -   An aliquot of the DTO soap was heated to 90° C., mixed with            finely ground SBP powder, and dried.    -   2. Saponified DTO

Gastrointestinal Digestion of the Saponified DTO:

Part of the liquid DTO soap and the carrier-absorbed DTO soap wasdigested by a pepsin-HCl-treatment (pH 2.25) followed by a pancreatinbile-acid-NaOH treatment (pH 6.2) in a dilution series. The digestionwas made to evaluate whether the products would resist the conditions ofthe upper gastrointestinal tract before they enter the distal intestinewith higher microbial activity.

The simulation was conducted in a total of 160 2-ml plasticmicrocentrifuge vials, in 1.5 ml volume, with 10 hours simulation time.Samples were tested at four concentrations of the dry matter of DTO: 0%,0.005%, 0.01%, 0.01% and 1%.

All the simulation samples were analysed for short chain fatty acids andthe total number of microbes. In addition, selected samples wereanalysed for a number of microbial species or groups by quantitativereal-time PCR (qPCR). Ileal simulation samples were analysed forlactobacilli and streptococci.

Results

In the ileal simulation model, DTO soap at 0.5 kg/ton level increasedthe concentrations of acetic and propionic acids and decreased theconcentration of lactic acid. This suggests modulation of microbialmetabolism from homofermentative towards heterofermentative metabolicalroute, which can be seen as a very positive change improving the feedconversion ratio. The sugar beet pulp carrier had little effect on thefermentation

Example 4 Test A: Toxin Adsorption into Solid Phase In Vitro

The capacity of a test product to remove toxins from aqueous medium wasmeasured in this test. An efficient toxin adsorbent should be able tobind the toxin in all compartments of the digestive tract, to inhibitthe toxin from getting absorbed by the animal. To evaluate the efficacyof the binder in the acidic stomach, the test was run at pH value 2.5(50 mM glycine-HCl buffer).

The test product was a saponified DTO product which contains 20% resinacids. The saponified DTO was manufactured as in example 3. The producttested was the saponified DTO (20%) with or without silicate carrier.

The test A was conducted with two toxins Ochratoxin A (OTA) andZearalenone (ZEA), at pH-value 2.5, three test substance levels 0.2, 0.5and 1 kg/ton and four replicate samples per treatment. Control treatmentwas replicated 8 times.

Mycotoxins OTA and ZEA were available as 3H-labeled pure compounds, andradioactivity, measured by liquid scintillation counting, was used fortheir quantification in the samples.

The experiment was conducted in silanized glass vials in 1 ml volume ofbuffer. In the test system, the bound radioactive toxin becomes removedfrom the liquid phase through co-pelleting with the insoluble componentsof the potential binder. The following procedure was used: 1. The testproducts were weighed into the vials, 2.3H-labeled and intact mycotoxinwas mixed with the buffers to get the final toxin concentration of 10μg/1, 3.1 ml of the buffermycotoxin solution was added to the vials, 4.The vials were sealed and kept for 2 hours at 37° C. in constant slowshaking, 5. The vials were centrifuged for 10 min at 3000×g 6.50 μl ofthe supernatant was mixed with 150 μl of liquid scintillation cocktail(Optiphase) into wells of a 96-well microtiter plate and 7. Theradioactivity of the samples was measured with a liquid scintillationcounter for five minutes

Results

The saponified DTO was able to bind OTA from the aqueous mediumstatistically significantly, and the binding was dependent on theconcentration of the test product. The saponified DTO adsorbed 25-60% ofthe free OTA from the medium.

The saponified DTO significantly decreased the amount of free ZEA evenat the lowest dosages. The saponified DTO removed approximately 30-60%of the free toxin.

It is obvious to a person skilled in the art that, with the advancementof technology, the basic idea of the invention may be implemented invarious ways. The invention and its embodiments are thus not limited tothe examples described above; instead they may vary within the scope ofthe claims.

The invention claimed is:
 1. A method for modulation of a microbialpopulation of an animal digestive tract, the method comprisingadministering to the animal an effective amount of a feed supplementcomprising tall oil rosin (TOR), wood rosin, GUM rosin, distilled talloil (DTO), or a mixture thereof, wherein the feed supplement comprises aresin acid based composition, the feed supplement contains over 12%(w/w) resin acids, and the feed supplement is in an amount of0.00001-1.0% (w/w) of a dry weight of a total amount of feed, andmodulating the microbial population.
 2. The method according to claim 1,further comprising improving feed utilization.
 3. The method accordingto claim 1, further comprising improving the feed conversion ratio. 4.The method according to claim 1, wherein the resin acid basedcomposition comprises at least one of the resin acids is selected fromthe group consisting of abietic acid, dehydoabietic acid, palustricacid, neoabietic acid, pimaric acid, isopimaric acid, and derivativethereof.
 5. The method according to claim 1, wherein the resin acidbased composition is Tall Oil Rosin (TOR).
 6. The method according toclaim 1, wherein the feed supplement comprises over 60% (w/w) of theresin acids.
 7. The method according to claim 1, wherein the resin acidbased composition is Wood Rosin.
 8. The method according to claim 1,wherein the resin acid based composition is GUM Rosin.
 9. The methodaccording to claim 1, wherein the feed supplement comprises over 10 andup to 99% (w/w) resin acids.
 10. The method according to claim 1,wherein the resin acid based composition is Distilled Tall Oil (DTO).11. The method according to claim 1, wherein the feed supplementcomprises over 10 and up to 60% (w/w) resin acids.
 12. The methodaccording to claim 1, wherein the resin acids are unmodified.
 13. Themethod according to claim 1, wherein the resin acids are modifiedchemically, biologically or other ways.
 14. The method according toclaim 13, wherein the resin acids are hydrogenated, disproportined,isomerizated, oxidized, polymerized, etherified, saponified and/oresterified and/or used as a reactant in Diels-Alder reaction.
 15. Themethod according to claim 1, wherein the feed supplement comprises 1-90(w/w) fatty acids and/or their derivatives.
 16. The method according toclaim 1, wherein the resin acids are modified by saponification.
 17. Themethod according to claim 1, wherein the resin acid based composition isdried.
 18. The method according to claim 1, wherein the resin acid basedcomposition is absorbed into a carrier material or mixed with a carrier.